Objective Rab11 GTPases play an essential role in regulating membrane trafficking pathways in eukaryotic cells. Nonetheless, there has been little work done on characterizing the transport machinery of Trichomonas. The aim of this study is to clone and characterize a Rab11 gene of Trichomonas vaginalis.Methods A cDNA expression library was constructed with T. vaginalis total RNA. A cDNA clone, which showed a high degree of homology with Rab proteins of different species, was isolated and sequenced. Sequence analysis was performed using BLASTP, RPS-BLAST and ClustalW programs. The genomic DNA corresponding to the cDNA sequence was amplified using PCR techniques and following by sequencing. Results cDNA with a length of 710 base pairs and an open reading frame of 636 bp was obtained. The deduced amino acid sequence from the open reading frame was found to possess 211 residuals. Sequence analysis demonstrated that this cDNA clone was homologous to the Rab11 subfamily of different species (60% identity and 79% similarity with Arabidopsis thaliana Rab11c, 58% identity and 78% similarity with human Rab11b), and that the amino acid sequence contains all the well known conserved sequence elements of Rab family. Specific Rab motifs were also detected in the deduced amino acid sequence. Phylogenetic analysis showed that its closest homologues are Rab11 proteins from other species. Sequencing of the PCR product of genomic DNA revealed that the genomic DNA sequence encompassing the putative 5'-ATG and 3'-stop codon is identical to the cDNA sequence.Conclusion A cDNA clone corresponding to the T. vaginalis Rab11 gene was obtained.The function of this gene in regulating membrane trafficking pathways of the parasitic protist is still under investigation.

Objective To summarize the operational experience and explore the best surgery strategy in cicatricial alopecia.Methods Clinical date of 80 cases of cicatricial alopecia during January 2012 to December 2016 were analyzed retrospectively.The operation methods and related adverse events were recorded.Patients were followed-up on the postoperative 1 week,1 month,3 months,6 months and 1-2 years.The outcomes were evaluated by a 4-levels questionnaire:very satisfied,satisfied,not satisfied,and no effect.Results Forty cases were operated with expanded skin flap + Follicular unit extraction (FUE) transplantation,10 cases with scar resection + FUE repair,and 30 cases only with FUE.Twenty cases were completed treatment with single-stage surgical operation,and 60 cases with two-stage surgical operation.A percentage (70％) of patients was very satisfied and 30％ were satisfied after one-stage surgical operation.A percentage (85％) of patients was very satisfied and 15％ were satisfied after two-stage surgical operation.Conclusions The cicatricial alopecia needs comprehensive surgical treatment.FUE is a best additional operation technology.The effect of combined treatment is better than single therapy method in large area cicatricial alopecia.

Objective To study and analyze the risk factors in children with type 1 diabetes and formulate preventive health measures. Methods A total of 112 children with type 1 diabetes treated in our hospital from January 2017 to October 2019 were selected as the type 1 diabetes group, and 50 healthy children who underwent physical examination during the same period were selected as the control group. Multifactor logistic regression analysis was used to analyze predisposing factors of type 1 diabetes in children, and preventive health measures was proposed. Results The results of multivariate logistic regression analysis indicated that maternal age, passive smoking during pregnancy, milk feeding time, and children's respiratory infections were independent risk factors for children with type 1 diabetes (OR: 6404, 6.903, 6.417, 8.256, P <0.05). Conclusion Maternal age, passive smoking during pregnancy, milk addition time, and children's respiratory infections were independent risk factors for children with type 1 diabetes. Strengthening health education, breastfeeding as soon as possible, and preventing respiratory infections can help reduce the incidence of children with type 1 diabetes.

OBJECTIVE：To investigate the effects of Biyuanshu （BYS）oral solution on IFN-γ of chronic rhinosinusitis （CRS）model mice ，and to investigate its potential mechanism on the basis of B 7-H1/PD-1 immune checkpoint. METHODS ：Male C57 mice were randomly divided into normal group ，sham operation group ，chemical medicine control group （clarithromycin，103 mg/kg），BYS low-dose ，medium-dose and high-dose groups （BYS oral solution ，3.1，6.2，12.4 mL/kg），with 20 mice in each group. Except for normal group without any treatment ，other mice were all open maxillary sinus ，sham operation group was not filled with sponge with bacteria ，while model group and administration groups were filled with sponge with bacteria to induce CRS model. Since 8th week after modeling ，normal group ，sham operation group and model group were given normal saline 0.2 mL intragastrically，administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 14 d. The nasal symptoms and general condition of mice were observed ，and the pathological changes of mice ’s nasal sinus mucosa were observed by HE staining ；qRT-PCR was used to measure the mRNA expression of IFN-γ，B7-H1 and PD- 1 in nasal sinus mucosa of mice. RESULTS：The normal group and sham operation group had no abnormal in nose ，and the epithelium and cilia of the nasal sinus mucosa were intact ；there was no significant difference f8y3j0127@163.com in the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 between 2 groups（P＞0.05）. In model group ，the mice were found to have runny n ose，frequent scratching and sneezing ，a small amount of yellow secretion in the nasal cavity ，and serious depilation ；the nasal sinus mucosa was seriously damaged ，cilia was exfoliated ，and the gland in the submucosa was hyperplasia ，lymphocyte infiltration was also found ；the relative mRNA expression of IFN-γ，B7-H1 and PD- 1 were significantly increased compared with normal group （P＜0.01）. Compared with model group，the nasal symptoms ，general condition and pathological changes of the nasal sinuses in each administration group were improved in varying degrees ；the relative mRNA expression of IFN-γ and B7-H1 in chemical medicine control group ，BYS medium-dose and high-dose groups ，as well as the relative mRNA expression of PD- 1 in administration groups were decreased significantly；above indexes of BYS medium-dose and high-dose groups were significantly lower than BYS low-dose group ，while relative mRNA expression of IFN-γ in BYS high-dose group were significantly higher than BYS medium-dose group. The relative mRNA expression of IFN-γ in BYS low-dose and medium-dose groups，the relative mRNA expression of B 7-H1 in BYS low-dose group，the relative mRNA expression of PD- 1 in BYS groups were significantly higher than chemical medicine control group ； mRNA expression of IFN-γ in BYS high-dose group was significantly higher than chemical medicine control group（P＜0.05 or P＜ 0.01）. Above indexes of BYS medium-dose group were similar to those of chemical medicine control group （P＞0.05）. CONCLUSIONS：BYS oral solution can improve chronic inflammation in nasal sinus mucosa of mice ，the mechanism of which may be associated with intervening mRNA overexpression of B 7-H1/PD-1 by inhibiting mRNA expression of IFN-γ.

Transcriptional regulation is crucial for regulated gene expression. Due to the complexity, it has been difficult to engineer eukaryotic transcription factor (TF) and promoter pairs. The few availabilities of eukaryotic TF and promotor pairs limit their practical use for clinical or industrial applications. Here, we report a de novo construction of synthetic inhibitory transcription factor and promoter pairs for mammalian transcriptional regulation. The design of synthetic TF was based on the fusion of DNA binding domain and Kruppel associated box transcription regulating domain (KRAB). The synthetic promoter was constructed by inserting the corresponding TF response element after SV40 promoter. We constructed and tested five synthetic inhibitory transcription factor and promoter pairs in cultured mammalian cells. The inhibition capability and orthogonality were verified by flow cytometry. In summary, we demonstrate the feasibility of constructing mammalian inhibitory TF and promoter pairs, which could be standardized for advanced gene-circuit design and various applications in the mammalian synthetic biology.
Animals ; Gene Expression Regulation ; Gene Regulatory Networks ; Mammals ; Promoter Regions, Genetic ; Transcription Factors ; Transcription, Genetic